The Utility of Nanopore Technology for Protein and Peptide Sensing

Abstract

Resistive pulse nanopore sensing enables label-free single-molecule analysis of a wide range of analytes. An increasing number of studies have demonstrated the feasibility and usefulness of nanopore sensing for protein and peptide characterization. Nanopores offer the potential to study a variety of protein-related phenomena that includes unfolding kinetics, differences in unfolding pathways, protein structure stability, and free-energy profiles of DNA-protein and RNA-protein binding. In addition to providing a tool for fundamental protein characterization, nanopores have also been used as highly selective protein detectors in various solution mixtures and conditions. This review highlights these and other developments in the area of nanopore-based protein and peptide detection.

Single‐Molecule Study of Peptides with the Same Amino Acid Composition but Different Sequences by Using an Aerolysin Nanopore

“…[27,28] The physical and chemical properties of the studied analyte including its size, concentration, conformation, structure, charge, and interaction with the pore can be inferred from statistical analysis of the blockade events. [29][30][31] The detection of peptides/proteins by the nanopore technique has become a research hotspot in recent years. Peptides or proteins are basic substances that make up cells, and play important roles in all aspects of life.…”

The applications of nanopores in studies of proteins

“…The need to better characterize protein properties and activities has led to the development of nanopores, which range from 1 to 100 nm in size and have the capacity to trace the ionic current passing through them upon application of a voltage [2]. Compared with other technologies, the advantages of nanopore-based methods for protein analysis are (1) their sensitivity to detect subtle changes in single proteins even at very low concentrations; (2) their ability to provide a dynamic view of protein conformation under physiological conditions; and (3) their label-free nature [2,13,14]. These advantages make nanopore technology a practical alternative and perhaps a more suitable approach for protein characterization.…”

“…To date, several review articles discussing the application of nanopores for protein sensing have been published [2,13,14]. For example, key nanopore materials, nanoscale features, and design requirements for detecting and characterizing proteins at the single-molecule level using nanopores have been described [2].…”

“…For example, key nanopore materials, nanoscale features, and design requirements for detecting and characterizing proteins at the single-molecule level using nanopores have been described [2]. Additionally, recent progress in nanopore-based studies of the translocation features of specific proteins bound to DNA [13] and the various means of identifying peptides, proteins, and their complexes [14] have been discussed. In the present review, we summarize and highlight several areas in the field of nanopore-based protein characterization, including studies of nanopore-based peptide detection and amino acid sequencing, measurement of single protein properties, dynamics of protein folding/unfolding and modifications, and interactions of proteins with other biomolecules (particularly DNA assembled nanostructures).…”

Polarization Induced Electro-Functionalization of Pore Walls: A Contactless Technology

“…Over the last twenty years, functionalized tridimensional pores have emerged as a specific range of biosensors offering sensitivities higher than those of conventional methods [1][2][3][4]. Pore sensing was successfully employed to detect and analyze different types of biomolecules and cells: single-and double-stranded nucleic acids [5], peptides [6], proteins [7], bacteria [8,9], viruses [10,11], and cancer cells [12,13]. Scheme 1 describes the principle of detection of a single biomolecule passing through a 2 of 19 pore: When crossing the pore, the target partially blocks the aperture (Scheme 1A), which is detected by a variation in the ionic current (Scheme 1B).…”